本文選題：藍(lán)光 + 抑制�。� 參考：《沈陽農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：果實(shí)的成熟過程是決定其貯藏性的重要因素。研究果實(shí)的成熟及調(diào)控機(jī)理對(duì)于人工調(diào)控果實(shí)貯藏性有重要的理論及實(shí)踐意義。秋子梨(Pyrus ussuriensis Maxim)是我國(guó)北方地區(qū)特有的梨資源,大部分秋子梨包括'南果梨'是呼吸躍變型果實(shí),其成熟過程主要受乙烯調(diào)控。除此之外,光、溫度等外界環(huán)境因素也可以調(diào)控果實(shí)的成熟過程。近年來許多研究證明了藍(lán)光能夠抑制呼吸躍變型果實(shí)成熟過程中的乙烯合成,從而影響果實(shí)的貯藏性,但其分子機(jī)制尚不清楚。本研究以'南果梨'果實(shí)為試材,研究了藍(lán)光通過其信號(hào)途徑中的轉(zhuǎn)錄因子PuHYH1抑制乙烯合成關(guān)鍵基因PuACS1的表達(dá)進(jìn)而調(diào)控果實(shí)成熟過程中的乙烯合成的分子機(jī)制。主要結(jié)果如下:1.從梨基因組克隆了光信號(hào)轉(zhuǎn)導(dǎo)途徑中的重要轉(zhuǎn)錄因子HY5的同源基因PuHYH1以及乙烯信號(hào)轉(zhuǎn)導(dǎo)途徑的重要轉(zhuǎn)錄因子ERF和乙烯合成的關(guān)鍵基因ACS、ACO,通過對(duì)其表達(dá)模式分析發(fā)現(xiàn),藍(lán)光處理后PuHYH1的表達(dá)上調(diào)乙烯合成關(guān)鍵基因PuACS1的表達(dá)下調(diào),PuEPF2表達(dá)上調(diào)。2.酵母單雜交、EMSA試驗(yàn)發(fā)現(xiàn)PuHYH1和PnERF2均能夠結(jié)合PuACS1的啟動(dòng)子;通過GUS報(bào)告基因活性測(cè)定試驗(yàn)發(fā)現(xiàn)PuHYH1和PuERF2均可負(fù)調(diào)控PuACS1基因的表達(dá)。3.通過酵母雙雜交試驗(yàn)、Pull-down試驗(yàn)發(fā)現(xiàn)了 PuHYH1與PuERF2之間存在蛋白互作關(guān)系。4.通過GUS報(bào)告基因活性測(cè)定試驗(yàn)明確了 PuHYH1與PuERF2蛋白之間的互作可以增強(qiáng)PuERF2對(duì)PuACS1的負(fù)調(diào)控作用。以上結(jié)果表明:PuHYH1、PuERF2可以直接負(fù)調(diào)控PuACS1的表達(dá),PuHYH1與PuERF2互作可以增強(qiáng)PuERF2對(duì)PuACS1的負(fù)調(diào)控,藍(lán)光可能通過以上兩條途徑抑制PuACS1的表達(dá)進(jìn)而抑制乙烯合成。本實(shí)驗(yàn)結(jié)果從分子水平闡明了藍(lán)光抑制'南果梨'乙烯合成的機(jī)制,對(duì)如何提高'南果梨'的貯藏性研究提供了重要的理論基礎(chǔ)。
[Abstract]:The ripening process of fruit is an important factor to determine its storage property. The study of fruit maturation and regulation mechanism has important theoretical and practical significance for artificial regulation of fruit storage. Pyrus ussuriensis Maximis is a unique pear resource in northern China. Most of the autumn pears, including 'Nanguo pear', are respiratory jump type fruits, and their ripening process is mainly regulated by ethylene. In addition, light, temperature and other external environmental factors can also regulate the fruit ripening process. In recent years, many studies have proved that blue light can inhibit ethylene synthesis in the ripening process of respiratory jump fruits, thus affecting the storage ability of fruits, but its molecular mechanism is not clear. In this study, the molecular mechanism of blue light inhibiting the expression of ethylene synthesis key gene PuACS1 and regulating ethylene synthesis during fruit ripening was studied using the fruit of 'Nanguo pear' as the experimental material, and the transcription factor PuHYH1 in its signaling pathway was used to inhibit the expression of the key gene of ethylene synthesis. The main results are as follows: 1. The homologous gene PuHYH1 of important transcription factor HY5 in light signal transduction pathway, the important transcription factor ERF of ethylene signal transduction pathway and the key gene of ethylene biosynthesis, ACSACO, were cloned from the genome of pear. The expression of PuHYH1 upregulated after blue light treatment down-regulated the expression of PuACS1, the key gene of ethylene synthesis, and upregulated the expression of PuEPF2. The results showed that both PuHYH1 and PnERF2 could bind to the promoter of PuACS1, and PuHYH1 and PuERF2 could negatively regulate the expression of PuACS1 gene by GUS reporter gene activity assay. The protein interaction between PuHYH1 and PuERF2 was found by the pull-down test of yeast two-hybrid. 4. GUS reporter gene activity assay showed that the interaction between PuHYH1 and PuERF2 protein could enhance the negative regulation of PuERF2 on PuACS1. These results suggest that the interaction of PuHYH1 and PuERF2 can directly and negatively regulate the expression of PuACS1. The interaction between PuHYH1 and PuERF2 can enhance the negative regulation of PuACS1 by PuERF2. Blue light may inhibit the expression of PuACS1 and inhibit ethylene synthesis through these two pathways. In this study, the mechanism of blue light inhibiting ethylene synthesis of 'Nanguo pear' was elucidated at molecular level, which provided an important theoretical basis for the study on how to improve the storage ability of 'Nanguo pear'.
【學(xué)位授予單位】：沈陽農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S661.2
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本文編號(hào)：1826477